Expression of human adiponectin gene in the baculovirus-insect cell expression system

Objective To investigate the expression of recombinant human adiponectin(ADPN) using baculovirus expression system in Sf 9 insect cell line.Methods PCR method was used to amplify human ADPN gene,and then the gene was inserted into plasmid pFastBacl.Recombinant plasmid was transformed into E.coli DH10Bac containing a shuttle vector,Bacmid.Recombinant shuttle vector Bacmid-ADPN was obtained by site-specific transposition and transfect Sf 9 cells.To identify the expressed product by SDS-PAGE、Western blot.Results Sf 9 cells infected by the recombinant baculovirus was observed for morphological changes.The Mw of recombinant protein was about 30 ku and Western blot proved that expressed protein could combine with ADPN polyclonal antibody.ConclusionHuman ADPN gene was successfully expressed in Sf 9 cells,and thus provided the material for studying its bioactivity and mechanism of action.